Therapeutic treatments using the direct application of antimicrobial metal compositions

ABSTRACT

Therapeutic treatments using the direct application of selected structures of antimicrobial metals in free-standing powder form, solution form and/or suspension form in therapeutically effective amounts. The selected structures of antimicrobial metals serve as an antimicrobial agent, an anti-inflammatory agent, an immuno modulator agent, an enzyme modulator agent, and/or an anti-tumor agent, for human and/or animal use.

REFERENCE TO PENDING PRIOR PATENT APPLICATIONS

This patent application:

(1) claims benefit of pending prior U.S. Provisional Patent ApplicationSer. No. 60/285,884, filed Apr. 23, 2001 by Robert E. Burrell et al. forTHERAPEUTIC TREATMENTS USING THE DIRECT APPLICATION OF NOBLE METALCOMPOSITIONS (Attorney's Docket No. WEST-1 PROV), which patentapplication is hereby incorporated herein by reference; and

(2) is a continuation-in-part of pending prior U.S. patent applicationSer. No. 09/840,637, filed Apr. 23, 2001 by Robert E. Burrell et al. forTREATMENT OF ACNE (Attorney's Docket No. 53-01), which patentapplication is also hereby incorporated herein by reference.

FIELD OF THE INVENTION

This invention relates to therapeutic treatments in general, and moreparticularly to therapeutic treatments using the direct application ofantimicrobial metal compositions.

BACKGROUND OF THE INVENTION

Localized infections affect millions of people each year. If not timelytreated, localized infections may spread, can result in unnecessarypain, may require increasingly more aggressive treatment, can result indevelopmental delays and permanent disability and, in severe cases, caneven result in death.

A common form of treatment for localized infections is oral antibiotictherapy. However, this treatment is systemic, requires multiple dosages,frequently causes side effects, and can give rise to the evolution ofantibiotic-resistant bacteria. Furthermore, debates are common amonghealthcare professionals and the general population regarding theover-use of antibiotics.

As a result, there is a significant need for an improved treatment forlocalized infections.

DESCRIPTION OF THE INVENTION Overview

Nucryst Pharmaceuticals Corp. and its predecessors, all of FortSaskatchewan, Alberta, Canada and sometimes collectively referred toherein as “Nucryst”, have developed selected structures of antimicrobialmetals such as silver, gold, platinum, palladium, etc. See, for example,International Patent Publication No. WO 93/23092, published Nov. 25,1993; International Patent Publication No. WO 95/13704, published May26, 1995; and International Patent Publication No. WO 98/41095,published Sep. 24, 1998, which documents are hereby incorporated hereinby reference. These selected structures of antimicrobial metals willhereinafter sometimes be collectively referred to as “antimicrobialmetals with atomic disorder”.

Nucryst has determined that its antimicrobial metals with atomicdisorder provide excellent therapeutic benefits. More particularly,antimicrobial metals with atomic disorder have been found to serveeffectively as an antimicrobial agent, an anti-inflammatory agent, animmuno modulator agent, an enzyme modulator agent, and/or an anti-tumoragent, for human and/or animal use. Among other things, antimicrobialmetals with atomic disorder have proven to be a broad spectrum (e.g.,gram positive, gram negative, fungus and drug resistant) bacteriocidalagent with little likelihood of fostering resistant bacteria and havinga sustained antimicrobial activity (for example, antimicrobial activitylasting over seven days has been consistently demonstrated with.antimicrobial metals with atomic disorder).

Furthermore, Nucryst has recently discovered new ways of forming itsantimicrobial metals with atomic disorder in free-standing powder form,solution form and suspension form. The ability to form antimicrobialmetals with atomic disorder in free-standing powder form, solution formand suspension form has greatly expanded the possibilities for usingthese selected nanocrystalline compositions for therapeutic purposes. Inparticular, the ability to form antimicrobial metals with atomicdisorder in free-standing powder form, solution form and suspension formhas lead to the further discovery that it is possible to utilizeantimicrobial metals with atomic disorder in a radical new way, i.e.,for direct application to a wide range of different tissues so as totherapeutically treat a wide range of different medical disorders.

Furthermore, by combining the therapeutic benefits of antimicrobialmetals with atomic disorder with recent advances in minimally invasivesurgery, including small-needle and needle-less drug delivery systems,therapeutic treatments may be applied to internal anatomy as well as tosurface anatomy.

Antimicrobial Metals With Atomic Disorder

The present invention utilizes selected structures of antimicrobialmetals. The antimicrobial metals are preferably selected noble metalssuch as silver, gold, platinum, palladium, etc. The structures areformed with atomic disorder, such that ions, clusters, atoms ormolecules of the metals are released at a concentration sufficient toprovide a localized therapeutic effect. The structures are preferably innanocrystalline form. Antimicrobial metals with atomic disorder may beprepared in the manner taught in International Patent Publication No. WO93/23092, published Nov. 25, 1993; International Patent Publication No.WO 95/13704, published May 26, 1995; and International PatentPublication No. WO 98/41095, published Sep. 24, 1998, which documentsare incorporated herein by reference.

As used herein, the terms and phrases set out below are intended to havethe meanings as follows:

“Metal” or “metals” includes one or more metals whether in the form ofsubstantially pure metals, alloys or compounds such as oxides, nitrides,borides, sulphides, halides or hydrides.

“Antimicrobial metals” are silver, gold, platinum, palladium, iridium,zinc, copper, tin, antimony, bismuth, or mixtures of these metals withsame or other metals, silver, gold, platinum and palladium beingpreferred, and silver being most preferred.

“Noble metals” are silver, gold, platinum and palladium, or mixtures ofsuch metals with same or other metals, with silver metal being the mostpreferred.

“Antimicrobial effect” means that atoms, ions, molecules or clusters ofthe antimicrobial or noble metal are released into the electrolyte whichthe coating contacts in concentration sufficient to inhibit microbialgrowth on and in the vicinity of the coating. The most common methods ofmeasuring an antimicrobial effect are a zone of inhibition test (whichindicates an inhibitory effect, whether microbiostatic or microbiocidal)or a logarithmic reduction test (which indicates a microbiocidaleffect). In a zone of inhibition test (ZOI) the material to be tested isplaced on a bacterial lawn (or a lawn of other microbial species) andincubated. A relatively small or no ZOI (ex. less than 1 mm) indicates anon-useful antimicrobial effect, while a larger ZOI (ex. greater than 5mm) indicates a highly useful antimicrobial effect. The ZOI is generallyreported as a corrected zone of inhibition (CZOI), wherein the size ofthe test sample is subtracted from the zone. A logarithmic reductiontest in viable bacteria is a quantitative measure of the efficacy of anantibacterial treatment; for example, a 5 log reduction means areduction in the number of microorganisms by 100,000-fold (e.g., if aproduct contained 100,000 pertinent microorganisms, a 5 log reductionwould reduce the number of pertinent microorganisms to 1). Generally, a3 log reduction represents a bactericidal effect. The logarithmicreduction test involves combining the inoculum with the test treatment,recovering the bacteria or other microbial species, and enumerating thebacteria or other microbial species using serial dilutions.

“Anti-inflammatory effect” means a reduction in one or more of thesymptoms of erythema (redness), edema (swelling), pain and prurituswhich are characteristic of inflammatory skin conditions.

“Inflammatory skin conditions” refers to those conditions of the skin inwhich inflammatory cells (e.g., polymorphonuclear neutrophils andlymphocytes) infiltrate the skin with no overt or known infectiousetiology, but excluding psoriasis and its related conditions. Symptomsof inflammatory skin conditions generally include erythema (redness),edema (swelling), pain, pruritus, increased surface temperature and lossof function. As used herein, inflammatory skin conditions include, butare not limited to, eczema and related conditions, insect bites,erythroderma, mycosis fungoides and related conditions, pyodermagangrenosum, erythema multiforme, rosacea, onychomycosis, and acne andrelated conditions, but excluding psoriasis and its related conditions.

“Biocompatible” means generating no significant undesirable hostresponse for the intended utility. Most preferably, biocompatiblematerials are non-toxic for the intended utility. Thus, for humanutility, biocompatible is most preferably non-toxic to humans or humantissues.

“Sustained release” or “sustainable basis” are used to define release ofatoms, molecules, ions or clusters of a antimicrobial metal thatcontinues over time measured in hours or days, and thus distinguishesrelease of such metal species from the bulk metal, which release suchspecies at a rate and concentration which is too low to betherapeutically effective, and from highly soluble salts ofantimicrobial metals such as silver nitrate, which releases silver ionsvirtually instantly, but not continuously, in contact with an alcohol orelectrolyte.

“Atomic disorder” includes high concentrations of one or more of: pointdefects in a crystal lattice, vacancies, line defects such asdislocations, interstitial atoms, amorphous regions, grain and sub grainboundaries and the like relative to its normal ordered crystallinestate. Atomic disorder leads to irregularities in surface topography andinhomogeneities in the structure on a nanometer scale.

“Normal ordered crystalline state” means the crystallinity normallyfound in bulk metal materials, alloys or compounds formed as cast,wrought or plated metal products. Such materials contain only lowconcentrations of such atomic defects as vacancies, grain boundaries anddislocations.

“Diffusion”, when used to describe conditions which limit diffusion inprocesses to create and retain atomic disorder, i.e. which freeze-inatomic disorder, means diffusion of atoms (adatom diffusion) and/ormolecules on the surface or in the matrix of the material being formed.

“Alcohol or water-based electrolyte” is meant to include any alcohol orwater-based electrolyte that the antimicrobial materials of the presentinvention might contact in order to activate (i.e. cause the release ofspecies of the antimicrobial metal) into same. The term is meant toinclude alcohols (short chain (C₆or less) and preferably C₄ or less),water, gels, fluids, solvents, and tissues containing, secreting, orexuding water or water-based electrolytes, including body fluids (forexample blood, urine, or saliva), and body tissue (for example skin).

“Bioabsorbable” as used herein in association includes substrates whichare useful in medical devices, that is which are biocompatible, andwhich are capable of bioabsorption in period of time ranging from hoursto years, depending on the particular application.

“Bioabsorption” means the disappearance of materials from their initialapplication site in the body (human or mammalian) with or withoutdegradation of the dispersed polymer molecules.

“Colour change” is meant to include changes of intensity of light undermonochromatic light as well as changes of hue from white lightcontaining more than one wavelength.

An “interference colour” is produced when light impinges on two or morepartly reflective surfaces separated by a distance which bears the rightrelationship to the wavelength of the light to be removed by destructiveinterference.

“Partly reflective” when used to describe the base or top layermaterials, means that the material has a surface which reflects aportion of incident light, but which also transmits a portion of theincident light. Reflection occurs when a ray of incoming lightencounters a boundary or interface characterized by a change inrefractive index between two media. For the top layer of theantimicrobial materials of this invention, that interface is with air.For the base layer, the interface is with the top layer. The reflectanceof the base and top layers is balanced so as to generate an interferencecolour.

“Partly light transmissive” when used to describe a thin film of the toplayer material means that the thin film is capable of transmitting atleast a portion of incident visible light through the thin film.

“Detectable” when used to describe a colour change means an observableshift in the dominant wavelength of the reflected light, whether thechange is detected by instrument, such as a spectrophotometer, or by thehuman eye. The dominant wavelength is the wavelength responsible for thecolour being observed.

“Cold working” as used herein indicates that the material has beenmechanically worked such as by milling, grinding, hammering, mortar andpestle or compressing, at temperatures lower than the recrystallizationtemperature of the material. This ensures that atomic disorder impartedthrough working is retained in the material.

“Pharmaceutically- or therapeutically-acceptable” is used herein todenote a substance which does not significantly interfere with theeffectiveness or the biological activity of the active ingredients(antimicrobial and anti-inflammatory activities) and which has anacceptable toxic profile for the host to which it is administered.

“Therapeutically effective amount” is used herein to denote any amountof a formulation of the antimicrobial or noble metals which will exhibiteither or both of an antimicrobial and optionally an anti-inflammatoryeffect, or some other therapeutic effect, when applied to the affectedarea of the tissue. A single application of the formulations of thepresent invention may be sufficient, or the formulations may be appliedrepeatedly over a period of time, such as several times a day for aperiod of days or weeks. The amount of the active ingredient, that isthe antimicrobial or noble metal in the form of a coating, powder ordissolved in liquid solution, will vary with the conditions beingtreated, the stage of advancement of the condition, the age and type ofhost, and the type and concentration of the formulation being applied.Appropriate amounts in any given instance will be readily apparent tothose skilled in the art or capable of determination by routineexperimentation.

“Carrier” means a suitable vehicle including one or more solid,semisolid or liquid diluents, excipients or encapsulating substanceswhich are suitable for administration to the skin.

“Nanocrystalline” is used herein to denote single-phase or multi-phasepolycrystals, the grain size of which is less than about 100, morepreferably <50, even more preferably <40, even more preferably <30, andmost preferably <25 nanometers in at least one dimension. The term, asapplied to the crystallite or grain size in the crystal lattice ofcoatings, powders or flakes of the antimicrobial or noble metals, is notmeant to restrict the particle size of the materials when used in apowder form.

“Powder” is used herein to include particulates of the antimicrobial ornoble metals ranging from nanocrystalline (less than 100 nm) tosubmicron sized powders up to flakes. Preferably, powders of theantimicrobial or noble metals used in the present invention are sized atless than 100 μm, and more preferably less than 40 μm, and mostpreferably less than 10 μm.

“Grain size”, or “crystallite size” means the size of the largestdimension of the crystals in the antimicrobial metal coating or powder.

“Hydrocolloid” means a synthetically prepared or naturally occurringpolymer capable of forming a thickened gel in the presence of water andpolyols (swelling agent). The swelling agent must be capable of swellingthe hydrocolloid chosen in order to form the gel phase.

“Hydrogels” means a hydrocolloid swollen with water or anotherhydrophilic liquid which is used for absorbing or retaining moisture orwater.

“Gel” means a composition that is of suitable viscosity for suchpurposes, e.g., a composition that is of a viscosity that enables it tobe applied and remain on the skin.

When used herein and in the claims, the term “nanocrystallineantimicrobial metal” and similar terminology, such as “nanocrystallinecoatings or powders” is meant to refer to antimicrobial metals formedwith atomic disorder and having a nanocrystalline grain size.

Free-Standing Powder Form Of Antimicrobial Metals With Atomic Disorder

Antimicrobial metals with atomic disorder may be provided infree-standing powder form in a variety of different ways.

By way of example but not limitation, in International PatentPublication No. WO 93/23092, published Nov. 25, 1993; and/orInternational Patent Publication No. WO 95/13704, published May 26,1995; and/or International Patent Publication No. WO 98/41095, publishedSep. 24, 1998, it was disclosed that antimicrobial metals with atomicdisorder may be provided in free-standing powder form by “cold working”.

Furthermore, in International Patent Publication No. WO 93/23092,published Nov. 25, 1993; and/or International Patent Publication No. WO95/13704, published May 26, 1995; and/or International PatentPublication No. WO 98/41095, published Sep. 24, 1998, it was disclosedthat antimicrobial metals with atomic disorder may be provided infree-standing powder form by vapor deposition on a fixed element, withthe deposited material thereafter being stripped off so as to yield thedesired free-standing powder.

In accordance with a further aspect of the present invention, it hasrecently been discovered that antimicrobial metals with atomic disordermay be provided in free-standing powder form by vapor deposition on anNGRC (“next generation roll coater”) apparatus. More particularly, acontinuous belt is coated with antimicrobial metals with atomic disorderusing vapor deposition techniques, with the antimicrobial metals withatomic disorder being scraped off the belt further down the line so asto yield the free-standing powder form of the antimicrobial metals withatomic disorder.

This last-mentioned method for making the free-standing powder form ofthe antimicrobial metals with atomic disorder (i.e., vapor deposition onan NGRC apparatus) is particularly advantageous, inasmuch as it canquickly and easily yield a relatively large supply of free-standingpowder at a relatively low cost.

Crystalline powder forms of the antimicrobial or noble metals(particularly preferred being Ag, Au, Pt, and Pd) can be prepared asfree standing powders, by coating powdered substrates, or from coatingson substrates which are then collected, for example by scraping and thensized. The powders may be prepared as pure metals, metal alloys orcompounds such as metal oxides or metal salts, by vapour deposition,mechanical working, or compressing to impart the atomic disorder. Thecrystalline powders are formed with atomic disorder in accordance withthe techniques set out above and as published in the prior patentapplications of Burrell et al., see for example WO 93/23092, publishedNov. 25, 1993, and WO 95/13704, published May 26, 1995. The atomicdisorder will most typically be formed in the metal powders duringphysical vapour deposition as set out above for coatings or bymechanically imparting the disorder, such as by milling, grinding,hammering, mortar and pestle or compressing, under conditions of lowtemperature (i.e., temperatures less than the temperature ofrecrystallization of the material) to ensure that annealing orrecyrstallization does not take place.

Alternatively, the powders may be formed by inert-gas condensationtechniques, which are modified to provide atomic disorder in the powderproduced, as taught in WO 95/13704 to Burrell et al.

Powders of the antimicrobial or noble metals are preferably formed byphysical vapour deposition (PVD) onto a substrate such as a cold finger,a silicon wafer, solid plates, a rotating cylinder, a continuous belt ina roll coater, or on steel collectors in known PVD coaters. Preparationof powders of the present invention by sputtering onto a continuous beltin a roll coater, or other some other moving or rotating substratesurface is particularly advantageous, inasmuch as it can quickly andeasily yield a relatively large supply of free-standing powder at arelatively low cost. A stainless-steel belt can be used in the rollcoating process without the need to provide additional cooling of thesubstrate. The powders or coatings are then scraped off to form apowder, and may be sized to avoid overly large particulates. The powdersare scraped off the moving surface with scrapers which contact themoving surface at an angle sufficient to remove the coating in flake orpowder form. The coating may be scraped off with scrapers angled forforward cutting of the coating from the moving surface, or with scraperswhich remove the coating from the moving surface by reverse draggingaction on the surface. The scrapers may be suspended above the belt, andeither weighted or spring loaded to apply pressure sufficient to removethe coating from the moving surface. With a continuous belt, thescrapers can conveniently be located above the end rollers to remove thecoating with a reverse dragging action as the belt rounds the endroller.

Alternatively, the powders of the antimicrobial or noble metals may beformed on powdered substrates which are biocompatible, or otherwisecompatible for the end use of the powder. Particularly preferredpowdered substrates are hydrocolloids, particularly those which arebioabsorbable and/or hygroscopic powders such as chitin. Exemplarybioabsorbable and/or hygroscopic powders are composed of:

Synthetic Bioabsorbable Polymers: for example polyesters/polyactonessuch as polymers of polyglycolic acid, glycolide, lactic acid, lactide,dioxanone, trimethylene carbonate etc., polyanhydrides, polyesteramides,polyortheoesters, polyphosphazenes, and copolymers of these and relatedpolymers or monomers.

Naturally Derived Polymers: Proteins: albumin, fibrin, collagen,elastin; Polysaccharides: chitosan, alginates, hyaluronic acid; andBiosynthetic Polyesters: 3-hydroxybutyrate polymers.

The powders may be incorporated into or onto medical dressings orpharmaceutical formulations, by any methods known in the art. Forexample, the powders may be layered onto the substrates (dressings orpowders), mechanically fixed within the fibres of the dressings,impregnated into dressings by physical blowing, or added to topicalpharmaceutical ingredients.

Preferably, powders of the present invention are sized at less than 100μm, and more preferably less than 40 μm, and most preferably about 3-5μm in size.

Once antimicrobial metals with atomic disorder have been provided infree-standing powder form, they can then be used therapeutically in thatform, or the free-standing powder can be used to form solutions orsuspensions of the antimicrobial metals with atomic disorder prior tobeing used to therapeutically treat tissue.

Use Of Antimicrobial Metals With Atomic Disorder In Free-standing PowderForm

Antimicrobial metals with atomic disorder in free-standing powder formmay be sprinkled lightly onto surface anatomy (e.g., the skin) intherapeutically effective amounts so as to provide an antimicrobialtreatment to that surface anatomy, e.g., to an infected cut. If desired,antimicrobial metals with atomic disorder may be mixed with one or moreother materials prior to being sprinkled onto the skin, where theseother materials may be biologically active materials (e.g., growthpromoters) or biologically neutral materials acting as a “filler” tofacilitate easier deployment of relatively small quantities ofantimicrobial metals with atomic disorder.

Or antimicrobial metals with atomic disorder in free-standing powderform can be applied to the lungs using a so-called dry powder inhaler.

Antimicrobial metals with atomic disorder in free-standing powder formmay also be injected, by small-needle or needle-less injection, into theinterior of the body in therapeutically effective amounts so as toprovide their therapeutic benefit to interior anatomy.

The antimicrobial metals with atomic disorder can be delivered tointerior anatomy via a small-needle drug delivery system or via aneedle-less drug delivery system. Such systems are available fromPowderject Research Limited of Oxford, United Kingdom (see, for example,U.S. Pat. Nos. 5,899,880; 6,010,478 and 6,013,050, which patents arehereby incorporated herein by reference) and Bioject, Inc. of Portland,Oreg. (see, for example, U.S. Pat. Nos. 4,596,556; 4,790,824; 5,064,413;5,312,335; 5,383,851; 5,399,163; 5,520,639; 6,096,002; and Des. 349,958,which patents are also hereby incorporated herein by reference).Delivery of antimicrobial metals with atomic disorder with such drugdelivery systems provides local therapy to the interior anatomy.

Where the antimicrobial metals with atomic disorder are to be applied tothe tissue in free-standing powder form by inhalation and/or injection,it is preferred that the particulate size be less than 2 microns, andpreferably less than 1 micron, so as so minimize any adverse reaction tothe presence of the particulate in the tissue.

Many health afflictions can be addressed by delivering antimicrobialmetals with atomic disorder, in free-standing powder form, to aninterior anatomical site with small-needle and/or needle-less drugdelivery systems. Examples of some of these applications include: (1)dermal drug delivery for skin conditions such as, but not limited to,acne, psoriasis, eczema and skin infections; (2) localized infectionssuch as, but not limited to, middle ear infections, endocardititis,pericarditis, prostatitis, sinusitis, osteomyelitis and onychomycosis;(3) mouth, gum and throat afflictions; (4) arthritis; and (5)direct-to-tumor chemotherapeutic delivery.

For example, antimicrobial metals with atomic disorder may be injecteddirectly into psoriatic plaques. Or antimicrobial metals with atomicdisorder may be injected by small-needle or needle-less injectionthrough the eardrum (i.e., through the tympanic membrane) into themiddle ear, whereby to provide a localized antimicrobial,anti-inflammatory treatment for middle ear infections. Or long-actingantimicrobial metals with atomic disorder may be injected into theprostate gland for difficult-to-treat prostate infections. Andantimicrobial metals with atomic disorder may be injected into thetissues of the oral cavity and throat to treat sore throats, thrush(candida infections) and periodontal diseases such as gingivitis. Also,antimicrobial metals with atomic disorder may be injected into arthriticjoints to reduce destructive inflammation. And long-acting antimicrobialmetals with atomic disorder of platinum can be injected into tumors thatare responsive to chemotherapy with platinum compounds.

Numerous advantages are achieved by delivering antimicrobial metals withatomic disorder to the interior anatomy using such drug deliverysystems. For one thing, local treatment (versus systemic treatment)results in lower total doses being required and in fewer side effects.For another thing, the broad spectrum antimicrobial activity of theantimicrobial metals with atomic disorder results in faster infectionfighting with low likelihood of bacterial resistance. Furthermore, dueto the long-acting nature of the antimicrobial metals with atomicdisorder, a single dose or relatively infrequent (e.g., weekly) dosesresults in an easier therapy regimen than many conventional treatmentregimens.

Solutions Of Antimicrobial Metals With Atomic Disorder

It is also possible to provide antimicrobial metals with atomic disorderin solution form. The solution form of antimicrobial metals with atomicdisorder can be advantageous in many anatomical applications, sincethere is substantially no particulate present which might irritatetissue.

Antimicrobial metals with atomic disorder may be provided in solutionform in a variety of different ways.

In one form of the invention, a solution of antimicrobial metals withatomic disorder is created by dissolving a free-standing powder ofantimicrobial metals with atomic disorder in water. The free-standingpowder of antimicrobial metals with atomic disorder may be packaged in a“tea-bag” type pouch, such that undissolved antimicrobial metals withatomic disorder remain captured within the pouch.

In another form of the invention, a solution of antimicrobial metalswith atomic disorder may be provided by immersing, in water, a substratecarrying deposited antimicrobial metals with atomic disorder. By way ofexample, a carrier strip may be coated with antimicrobial metals withatomic disorder by vapor deposition, and then the carrier strip may beimmersed in water so as to create the solution of antimicrobial metalswith atomic disorder. Alternatively, a bandage may be coated withantimicrobial metals with atomic disorder by vapor deposition, and thenthe bandage may be immersed in water so as to create the solution ofantimicrobial metals with atomic disorder.

The solution of antimicrobial metals with atomic disorder may beprepared in advance. (e.g., at a manufacturing plant) or on site at thetime of use. Where a solution of antimicrobial metals with atomicdisorder is prepared in advance (e.g., at a manufacturing plant), it ispreferred that the solution be created by immersing a “tea-bag” typepouch of the free-standing powder form of antimicrobial metals withatomic disorder in water and leaving it there until the time of use, orby immersing a substrate carrying deposited antimicrobial metals withatomic disorder in water and leaving it there until the time of use.

Once a solution of antimicrobial metals with atomic disorder has beencreated, it may be applied to tissue as a liquid or as an aerosol.

Regardless of how the solution of antimicrobial metals with atomicdisorder is applied to tissue, the. dosage is dependent, to at leastsome extent, on the concentration of antimicrobial metals with atomicdisorder present in the solution. Thus, where it is necessary to apply astrong dose of antimicrobial metals with atomic disorder, it may bedesirable to raise the concentration of antimicrobial metals with atomicdisorder in the solution. In this respect it has been discovered that bylowering the pH of the solution, a higher concentration of antimicrobialmetals with atomic disorder can be obtained and, significantly, theantimicrobial metals with atomic disorder go into solution faster. ThepH of the solution can be lowered by adding acid to the solution. In onepreferred form of the invention, CO₂ is added to the solution: the CO₂creates carbonic acid, thus lowering the pH of the solution andincreasing the concentration of antimicrobial metals with atomicdisorder in the solution.

As noted above, once a solution of antimicrobial metals with atomicdisorder has been created, it may be applied to tissue as a liquid or asan aerosol.

Use Of Antimicrobial Metals With Atomic Disorder In Solution Form—LiquidApplication

A solution of antimicrobial metals with atomic disorder may be applied,in liquid form, and in various viscosities, to a wide range of differenttissues in therapeutically effective amounts so as to therapeuticallytreat a wide range of different medical disorders.

By way of example, a solution of antimicrobial metals with atomicdisorder can be applied as a rinse or bath or wash to treat a dermalcondition such as, but not limited to, acne, psoriasis, eczema and skininfections. Alternatively, a solution of antimicrobial metals withatomic disorder can be applied as a rinse or bath or wash to treat awound or a surgical site.

Or a solution of antimicrobial metals with atomic disorder can beapplied to mouth tissue (e.g., the gums) as an oral rinse.

Or a solution of antimicrobial metals with atomic disorder can beapplied to throat tissue as a gargle.

Or a solution of antimicrobial metals with atomic disorder can beapplied to nasal passages and the sinus, e.g., to treat sinusitis andallergic rhinitis.

Or a solution of antimicrobial metals with atomic disorder can beapplied to the eyes as eyedrops.

Or a solution of antimicrobial metals with atomic disorder can beapplied to the ears as ear drops.

It is also possible to apply a solution of antimicrobial metals withatomic disorder, in liquid form, to internal anatomy using asmall-needle and/or needle-less drug delivery systems, includingcatheter-based drug delivery systems. Thus, for example, a solution ofantimicrobial metals with atomic disorder may be introduced by catheterinto the bladder to treat a bladder infection; or injected into themiddle ear to treat middle ear infections; or injected or instilled orotherwise introduced into the abdomen to treat a post-surgical abdominalabscess or to treat an infection from peritoneal dialysis; or injectedor instilled or otherwise introduced into other internal anatomicalstructures, including body cavities, so as to treat conditions such as,but not limited to, endocardititis, pericarditis, prostatitis,sinusitis, osteomyelitis and onychomycosis; or injected into skin tissueto treat acne, psoriasis, eczema and/or or other skin conditions; etc.

Use Of Antimicrobial Metals With Atomic Disorder In SolutionForm—Aerosol Application

A solution of antimicrobial metals with atomic disorder may also beapplied, in aerosol form, to a wide range of different tissues intherapeutically effective amounts so as to therapeutically treat a widerange of different medical disorders.

By way of example, a solution of antimicrobial metals with atomicdisorder may be applied in aerosol form to surface tissues as a spray.Thus, a solution of antimicrobial metals with atomic disorder can beapplied as a spray to treat a dermal condition such as, but not limitedto, acne, psoriasis, eczema and skin infections. Alternatively, asolution of antimicrobial metals with atomic disorder can be applied asa spray to treat or clean a wound or a surgical site.

By way of further example, a solution of antimicrobial metals withatomic disorder, in aerosol form, may be inhaled by a patient fordeployment to the throat, the nasal and sinus passages and/or the lungs.

The aerosol of antimicrobial metals with atomic disorder may be createdby passing a liquid solution of antimicrobial metals with atomicdisorder through a mechanical mister (e.g., a simple spray bottle ornebulizer) and may be applied directly (e.g., via a hand inhaler) orthrough some other delivery system (e.g., an oxygen tent, etc.).

With respect to an aerosol of antimicrobial metals with atomic disorder,it should be appreciated that the droplet size of the aerosol can beimportant for at least two reasons.

First, the droplet size of the aerosol can affect the dosage ofantimicrobial metals with atomic disorder being applied to the tissue,i.e., a larger droplet size results in delivery of more antimicrobialmetals with atomic disorder to the tissue.

Second, the droplet size of the aerosol can also affect delivery of theantimicrobial metals with atomic disorder to the target tissue, e.g.,where the aerosol is inhaled through the mouth, big droplets tend tostay in the throat whereas small droplets (e.g., approximately 10microns or so) tend to travel to the lungs.

Thus, depending on the dosage required and the. target tissue, it may beimportant to regulate the droplet size of the aerosol.

In this respect, it has been found that droplet size can be regulated,to at least some extent, by the device (e.g., the mechanical mister)which is used to produce the aerosol.

In addition, it has also been discovered that the aerosol's droplet sizecan be adjusted, to at least some extent, by modifying the surfacetension of the solution. More particularly, the solution ofantimicrobial metals with atomic disorder has water as its solvent, andwater has a relatively high surface tension, so it is relativelystraightforward to create an aerosol having a relatively small dropletsize. In accordance with the present invention, it has also beendiscovered that surfactants can be added to the solution so as to reducethe surface tension of the solution, whereby to create an aerosol havinga relatively large droplet size. By way of example, such surfactants maycomprise phospholipids, e.g., lecithin, sphingomyelin, etc.

Suspensions Of Antimicrobial Metals With Atomic Disorder

It is also possible to provide antimicrobial metals with atomic disorderin suspension form. The suspension form of antimicrobial metals withatomic disorder can be advantageous in many applications, since it has arelatively long storage life and, perhaps even more importantly, has arelatively long-lasting therapeutic life.

Antimicrobial metals with atomic disorder may be provided in suspensionform in a variety of different embodiments. More particularly, asuspension of antimicrobial metals with atomic disorder can be createdin free-standing form or as a dried gel applied to a medical device.

Thus, in one form of the invention, a suspension of antimicrobial metalswith atomic disorder can comprise a free-standing form, i.e., it cancomprise a liquid such as a lotion; or a semi-solid such as a gel (i.e.a water-based hydrocolloid) or an emulsion (i.e., an oil-in-water orwater-in-oil suspension) such as a cream or ointment. Formulations caninclude carboxymethyl cellulose (“CMC”), polyvinyl alcohol, methylparabin, proply parabin, and 0.1% antimicrobial metals with atomicdisorder in powder form.

In another form of the invention, a suspension of antimicrobial metalswith atomic disorder can comprise a dried gel applied to a medicaldevice. In this embodiment, a hydrated form of the gel is created,applied to a medical device, and then dehydrated. During use, the gelbecomes rehydrated, whereby the antimicrobial metals with atomicdisorder are released to provide their therapeutic effect to tissue.

Regardless of whether the suspension is free-standing or a dried gelapplied to a medical device, the suspension may also includebiologically active agents such as cytoconductive agents, etc. By way ofexample, betaglucan, a complex carbohydrate which appears to havecytoconductive properties, may be added to the suspension.

Of course, when formulating the suspension, care must be taken to avoidgenerating a blend which might deactivate the therapeutic effect of theantimicrobial metals with atomic disorder. Thus, for example, glycerolcan be deleterious to the therapeutic effect of the antimicrobial metalswith atomic disorder, and should be avoided.

Use Of Antimicrobial Metals With Atomic Disorder In SuspensionForm—Free-Standing Form

As noted above, a suspension of antimicrobial metals with atomicdisorder can comprise a free-standing form, i.e., it can comprise aliquid such as a lotion; or a semi-solid such as a gel (i.e. awater-based hydrocolloid) or an emulsion (i.e., an oil-in-water orwater-in-oil suspension) such as a cream or ointment. Thesefree-standing forms of the suspension are intended to be appliedtopically to the tissue which is to be treated, in therapeuticallyeffective amounts, and can be used to treat a dermal condition such as,but not limited to, acne, psoriasis, eczema and skin infections.Alternatively, the free-standing forms of the suspension can be appliedtopically to treat a wound or a surgical site, etc.

Use Of Antimicrobial Metals With Atomic Disorder In SuspensionForm—Dried Gel

It is also possible to provide a suspension of antimicrobial metals withatomic disorder in the form of a dried gel applied to medical devices.The hydrated form of the gel is created, applied to a medical device(e.g., during manufacture of the medical device), and then dehydrated.During use, the gel becomes rehydrated, whereby the antimicrobial metalswith atomic disorder are released in therapeutically effective amountsso as to provide their therapeutic effect to tissue.

Examples of medical devices which are prime candidates for a dried gelcoating include catheters (e.g., urological catheters, in-dwellingcatheters, drainage catheters, etc.), bone screws, total joints,vascular grafts, hernia meshes, surgical dressings, surgical packingmaterials, etc.

In this respect it should be appreciated that the dried gel can be quitestable and easy to handle when dehydrated, but very slippery whenrehydrated. Thus, dried gels can be particularly advantageous withcertain types of medical devices which might otherwise requirelubrication during use, since the rehydrated gel automatically providessuch lubrication. Urological catheters are one example of a medicaldevice which generally requires lubrication during use, and which wouldbenefit from the natural lubrication provided by the rehydrated gel.

It is also possible to provide a dried gel which is less slippery, oreven non-slippery, when rehydrated.

EXAMPLE 1

6 milligrams of antimicrobial metals with atomic disorder, infree-standing powder form, are sprinkled lightly onto 6.5 cm² of burnedtissue, and thereafter wet with a light spray of water or wound exudateor TDWL (Trans Dermal Water Loss) or other bodily fluids, so as toprovide an antimicrobial treatment to the burned tissue. The treatmentis repeated every 24 hours until the therapeutic effects are no longerneeded.

EXAMPLE 2

0.5 milligrams of antimicrobial metals with atomic disorder, infree-standing powder form, are injected, using a small-needle drugdelivery system or a needle-less drug delivery system, into gum tissueso as to treat gingivitis. The treatment is repeated every 3 days untilthe therapeutic effects are no longer needed.

EXAMPLE 3

A solution of antimicrobial metals with atomic disorder is prepared bydissolving 6 milligrams of antimicrobial metals with atomic disorder in1 gram of water. The solution of antimicrobial metals with atomicdisorder is applied as a rinse or bath or wash to a wound site so as toprovide an antimicrobial treatment to the wound site. The treatment isrepeated every 24 hours until the therapeutic effects are no longerneeded.

EXAMPLE 4

A solution of antimicrobial metals with atomic disorder is prepared bydissolving 6 milligrams of antimicrobial metals with atomic disorder in1 gram of water. The solution of antimicrobial metals with atomicdisorder is applied to the interior of the bladder via a catheter so asto provide antimicrobial treatment to the bladder. The treatment isrepeated every 8 hours until the therapeutic effects are no longerneeded.

EXAMPLE 5

A solution of antimicrobial metals with atomic disorder is prepared bydissolving 6 milligrams of antimicrobial metals with atomic disorder in1 gram of water. The solution of antimicrobial metals with atomicdisorder is injected (using a small-needle or needle-less injectionsystem) under the toenails or into the nail bed and/or the surroundingtissue of a person suffering from onychomycosis so as to provide anantimicrobial treatment to the tissue. The treatment is repeated 2 timesa day until the therapeutic effects are no longer needed.

EXAMPLE 6 Summary

Solutions of nanocrystalline noble metals were prepared by immersingActicoat® burn dressings (distributed by Smith & Nephew) in reverseosmosis water that had been pretreated with CO₂ in order to reduce thepH. Two different concentrations of antimicrobial metals with atomicdisorder solutions were prepared by this method, the concentrationsbeing 85 μg/mL and 318 μg/mL. Solutions of silver nitrate were alsoprepared to use as comparisons in the experiments. The concentrations ofthe silver nitrate were 103 ppm of silver and 295 ppm of silver asdetermined by Atomic Absorption Spectroscopy.

The solutions were in turn placed in an ultrasonic nebulizer thatcreated small droplets containing dissolved and suspended parts of thesolution of nanocrystalline noble metals. The output from the nebulizerwas directed into a chamber made from a stainless steel frame and base.Petri dishes containing Mueller Hinton agar streaked with 4 h oldcultures of Pseudomonas aerugiona and Staphylococcus aureus were exposedto nanocrystalline noble metal aerosols and the silver nitrate aerosols.

The results of the tests show that nanocrystalline noble metal aerosolstransmit the antimicrobial activity of the dressings to remote sites,and nanocrystalline noble metal aerosols are more effective thancomparable concentrations of silver nitrate.

Introduction

In many instances the delivery of antimicrobial materials may mostexpeditiously be accomplished by using aerosols (e.g., in the treatmentof pneumonia). The drawback of aerosols is the requirement for a highconcentration of the active ingredient to ensure that a sufficientamount is delivered to achieve the biological effect desired withoutcausing problems with the carrier solvent (e.g., water). The essentialrequirement of the equipment for producing an aerosol that containsdissolved and suspended components of antimicrobial metals with atomicdisorder is that it must form droplets of aerosol directly from theliquid form, and the aerosol droplets must be small enough to reach thelungs. This means that the droplets should be preferably less thanapproximately 10 μm. To meet these requirements, the aerosol cannot becreated by first evaporating the liquid and then condensing it to formdroplets, since this would remove the desired antimicrobial metals withatomic disorder from the aerosol. There are two methods that can be usedto relatively easily form the droplets directly: (1) mechanicaldisruption of the liquid; and (2) air, under pressure, passing throughsome form of orifice that combines the air and the liquid in a way thatcreates droplets instead of evaporating the liquid.

Several experiments were carried out with antimicrobial metals withatomic disorder and silver nitrate solutions to determine if theantimicrobial activity of the dressing could be transferred through adirect droplet aerosol to a Petri dish.

Equipment

The method used to create an aerosol for these tests was the mechanicalmethod in the form of an ultrasonic nebulizer. For convenience, anultrasonic humidifier was used. The liquid containing the dissolved andsuspended antimicrobial metals with atomic disorder was placed in thewater reservoir of the humidifier. When power was applied to thehumidifier, aerosol droplets of dissolved and suspended antimicrobialmetals with atomic disorder were generated and flowed from the outputnozzle.

A test chamber was constructed using a stainless steel frame with atransparent plastic covering. The frame was placed on a stainless steelplate. The output nozzle from the humidifier was modified so that theaerosol could be directed into the chamber at a height of approximately30 cm from the base. The plates and other test samples were placed onthe stainless steel plate and exposed to the aerosol for a prescribedlength of time.

Solution 1

A solution of antimicrobial metals with atomic disorder was prepared byimmersing 518 sq. inches of Acticoat® burn dressing in 1 L of reverseosmosis water, which was treated with CO₂ to maintain a pH of 6.5. After20 minutes the concentration of silver in the water was 85 μg/mL.

Solution 2

A solution containing 370 μg/mL of silver from a Acticoat® dressing wasprepared as follows: 1 L of reverse osmosis water was purged withcommercial grade carbon dioxide until the pH was 4.3. SufficientActicoat® dressing was added to bring the pH up to 6.5. At that time,the silver concentration was 370 μg/mL.

Solution 3

Ag as AgNo₃ was prepared by dissolving 0.157 g of AgNo₃ into 1 L ofreverse osmosis water and mixed until dissolved. The solution wasanalyzed by Atomic Absorption Spectroscopy and found to be 102.9 ppm ofsilver.

Solution 4

Ag as AgNO₃ was prepared by dissolving 0.427 of AgNO₃ into 1 L ofreverse osmosis water and mixed until dissolved. The solution wasanalyzed by Atomic Absorption Spectroscopy and found to be 295 ppm ofsilver.

Aerosolization

Petri dishes, containing Mueller Hinton agar, were streaked with 4 h oldcultures of Pseudomonas aeruginosa or Staphylococcus aureus. The plateswere then weighed and their exposed outer surfaces were coated withParafilm to prevent condensation from occurring on these surfaces. Theseplates were placed in the aerosol chamber uncovered. The ultrasonicnebulizer was then started and run for 53 minutes. The plates were thenremoved from the chamber, the plastic was removed and the dishesre-weighed so that the amount of moisture loss/gain could be determined.

The plates were then placed in a 35° C. incubator for 16 h. Afterincubation the pattern and amount of growth was assessed on the platesfor both organisms.

Viability Assessment

Three of the six plates made for each organism were tested to determineif the antimicrobial effect was cidal or static in nature. This wasaccomplished by rinsing or placing a piece of the clear section of agarin the Petri dish plates into Tryptic soy broth in a test tube andincubating for 4 h or 16 h. If the medium turned turbid in 4 h it wouldindicate that the antimicrobial affect was bacteriostatic in nature. Ifthe organism took more than 16 h to grow, as indicated by turbidity, itwas considered an indication that both static and cidal effectsoccurred. If no growth occurred, the effect was bactericidal.

Results

The results for Solutions 1 and 2 are summarized in Tables 1 and 2,respectively. TABLE 1 Solutions 1 and 3 Results Antimicrobial MetalsWith Atomic Disorder AgNo₃ Organism Ps. S. Ps. S. Aeruginosa aureusAeruginosa aureus Ag concentration 85 85 99 99 (μg/mL) pH of test 6.56.5 Approx. Approx. solution 6.5 6.5 Exposure time 53 53 53 53 (minutes)Exposed area 9.8 9.8 9.8 9.8 (sq. in) Weight gain (g) 0.8 0.8 1.05 1.05Growth at 16 h 0 0 0 ++++ (0-++++) at 48 h 0 ++ 0 ++++ Viable 4 h No YesNo Yes 16 h Yes Yes Yes Yes

TABLE 2 Solutions 2 and 4 Results Antimicrobial Metals With AtomicDisorder AgNo₃ Organism Ps. S. Ps. S. aeruginosa aureus aeruginosaaureus Ag concentration 370 370 300 300 (μg/mL) pH of test 6.5 6.5Approx. Approx. solution 6.3 6.3 Exposure time 53 53 53 53 (minutes)Exposed area 9.8 9.8 9.8 9.8 (sq. in) Weight gain (g) 1.14 1.14 1.121.12 Growth at 16 h 0 0 0 0 (0-++++) at 48 h 0 0 0 +++ Viable 4 h No NoNo No 16 h No No No N/A

Discussion

At the low concentration of silver in solution, the Acticoat® dressinggenerated silver was effective at controlling the growth of bothorganisms while the silver nitrate only prevented the growth of Ps.aeruginosa. Viability tests showed that at the low concentration,neither form of silver was completely bactericidal although the poorgrowth on the plates treated with antimicrobial metals with atomicdisorder compared to the silver nitrate treated plates suggests that asignificant log reduction occurred in the plates treated with theaerosol of antimicrobial metals with atomic disorder.

At a higher concentration of silver, both antimicrobial metals withatomic disorder (37.0 μg/mL) and AgNo₃ (300 μg/mL) were effective atcontrolling P. aeruginosa. Since no re-growth occurred, it is assumedthat the agent at this concentration was bactericidal. Antimicrobialsilver with atomic disorder was more effective than AgNO₃ at controllingS. aureus. No re-growth occurred on any plates or in the brothindicating a total kill of the organism while, in the AgNO₃ treatment, alarge number of organisms grew at 16h.

Based on weight gain during aerosol treatments, a dose per unit area canbe calculated. In each case for Solution 1, the dose was 8.5 μg/sq.inch, while for Solution 2, the dose was 38 μg/sq. inch. These doses, ona per lung basis, would be less than 10 mg of silver per hour oftreatment. Each hour of treatment with antimicrobial silver with atomicdisorder aerosols appears to provide at least 48 h of protection.Therefore, the dose per day, from the high concentration treatment,would be about 5 mg or a little less than the silver released by 2 sq.inches of SSD per day.

The most significant advantage of using antimicrobial silver with atomicdisorder may be the lack of a toxic action such as NO₃ or sulfadiazine.

CONCLUSIONS

(1) Aerosols of antimicrobial metals with atomic disorder transmit theantimicrobial activity of the dressings to remote sites.

(2) Aerosols of antimicrobial metals with atomic disorder are moreeffective than comparable concentrations of silver nitrate.

(3) The dose delivered is acceptable and would not appear to beexcessive.

(4) No toxic cations (NO₃ or sulfadiazine) are introduced to thepatient.

EXAMPLE 7 Gels of Antimicrobial Metals With Atomic Disorder

Gel products of antimicrobial metals with atomic disorder encompass both“wet” and “dry” materials.

A “wet” gel product of antimicrobial metals with atomic disorder is aproduct that provides moisture to a dry skin condition (psoriasis,eczema, acne, wound, etc.) and facilitates autolytic debridement ofnecrotic tissue. It also delivers the antimicrobial andanti-inflammatory properties of the suspended antimicrobial metals withatomic disorder powders.

In many instances it is also beneficial to supply biologically activemolecules to elicit a specific response such as cell migration, etc.Since these biologically active molecules are susceptible to microbialdegradation if not protected, it is beneficial to include them in gelsof antimicrobial metals with atomic disorder that will provide thenecessary protection.

“Dry” gel products of antimicrobial metals with atomic disorder arephysically stabilized (dry or cross-linked) materials that providelubricious, antimicrobial, antithrombogenic, and anti-inflammatoryproperties to a variety of implantable, trancutaneous or topicallyapplied devices. The coatings may also provide other benefits such asaccelerating or otherwise facilitating tissue integration by creating afavorable environment for cell proliferation. This favorable environmentmay be created by including cytoconductive agents or anti-adhesionagents such as bone morphogenetic proteins, B-glucan hyaluronic acids inthe gel. The gel may be stabilized by cross-linking the gel components(collagen, gelatin, etc.) or by drying the coated materials.

Examples of the primary gelling agents are listed in Table 3.Biologically active ingredients that may be used, in any combinationwith the primary gelling agents, are given in Table 4. Materials thatshould not be used with gels of antimicrobial silver with atomicdisorder are given in Table 5. TABLE 3 Material Percentage CompositionCarboxymethyl cellulose (CMC) 0.1-10 Polyvinyl alcohol (PVA) 0.1-10Collagen 0.1-10 Pectin 0.1-10 Gelatin 0.1-10 Chitin 0.1-10 Chitosan0.1-10 Alginate 0.1-10 Poly (α-amino acids) PolyesterPoly-1-caprolactone PEG Cocoa butter Sepigel

TABLE 4 Biologically Active Ingredients Percentage Composition Methylparaben <3 Propyl paraben <3 B-glucan <5 Hyaluronic acid <5 Epidermalgrowth factor <1 Platelet derived growth factor <1 Transforming growthfactor <1 Vascular endothelial growth factor <1 Interleukins <1 Heparin<5 Bone morphogenetic proteins <1

TABLE 5 Non-Compatible Materials Percentage Composition Chloridesalts >0.01 Aldehydes >0.01 Ketones >0.01 Long chain alcohols >0.01Glycerol >0.01 Triethanolamine >0.01

EXAMPLE 8 Examples of Gels with Antimicrobial Metals With AtomicDisorder No. 1

A commercial carboxymethyl cellulose/pectin gel (Duoderm Convatec) wascombined with antimicrobial metals with atomic disorder powder toproduce a gel with 0.1% silver. A logarithmic reduction test wasperformed as follows in the gel using Pseudomonas aeruginosa.

The inoculum was prepared by placing 1 bacteriologic loopful of theorganism in 5 mL of trypticase soy broth and incubating it for 3-4 h.The inoculum (0.1 mL) was then added to 0.1 mL of gel and vortexed(triplicate samples). The mixture was incubated for one-half hour. Then1.8 mL of sodium thioglycollate-saline (STS) solution was added to thetest tube and vortexed. Serial dilutions were prepared on 10⁻¹ to 10⁻⁷.A 0.1 mL aliquot of each dilution was plated in duplicate into Petriplates containing Mueller-Hinton agar. The plates were incubated for 48h and then colonies were counted. Surviving members of organisms weredetermined and the logarithmic reduction compared to the initialinoculum was calculated.

The logarithmic reduction for this mixture was 6.2, indicating asignificant bactericidal effect.

No. 2

Carboxymethyl cellulose (CMC) fibers were coated directly to produce adefective nanocrystalline antimicrobial coating. The CMC was then gelledin water by adding 2.9 g to 100 mL volume. This material was testedusing the method of No. 1. The material generated a 5.2 logarithmicreduction of Pseudomonas aeruginosa, demonstrating that the gel had asignificant bactericidal effect.

No. 3

An alginate fibrous substrate was directly coated with a defectivenanocrystalline antimicrobial coating. The alginate (5.7 g) was added to100 mL volume of water to create a gel. This material was tested usingthe method of No. 1. The material generated a 5.2 logarithmic reductionof Pseudomonas aeruginosa, demonstrating that the gel had a significantbactericidal effect.

No. 4

A commercial gel containing CMC and alginate (Purilin gel Coloplast) wasmixed with a defective nanocrystalline silver powder to give a productwith 0.1% silver. This was tested as above with both Pseudomonasaeruginosa and Staphylococcus aureus. Zone of inhibition data was alsogenerated for this gel as follows. An inoculum (Pseudomonas aeruginosaand Staphylococcus aureus) was prepared as in No. 1 and 0.1 mL of thiswas spread onto the surface of Mueller-Hinton agar in a Petri dish. Asix mm hole was then cut into the agar at the center of the Petri dishand removed. The well was filled with either 0.1 mL of the silvercontaining gel, a mupirocin containing cream or a mupirocin containingointment. The Petri plates were then incubated for 24 h and the diameterof the zone of inhibition was measured and recorded.

The silver containing gel produced 9 mm zones against both Pseudomonasaeruginosa and Staphylococcus aureus, while the mupirocin cream andointment produced 42 and 48 mm zones against Staphylococcus aureus and 0mm zones against Pseudomonas aeruginosa.

The silver containing gel reduced the Pseudomonas aeruginosa andStaphylococcus aureus properties 4.4 and 0.6 logs, respectively, showinggood bactericidal activity. The mupirocin cream and ointment generated0.4 and 0.8, and 0.8 and 1.6, log reductions against Staphylococcusaureus and Pseudomonas aeruginosa, respectively. The silver gel had botha greater bactericidal effect and spectrum of activity than themupirocin containing products.

Nos. 5-10

The formula for Nos. 5-10 are summarized in Table 6. Zones ofinhibitions were determined in No. 4 and log reductions were determinedin No. 1.

All formulae provided a broader spectrum of activity and a greaterbactericidal effect than did mupirocin in a cream or ointment form. Themupirocin cream produced zones of inhibition of 42 and 0, and logreduction of 0.4 and 0.8, against Staphylococcus aureus and Pseudomonasaeruginosa, respectively. TABLE 6 Antimicrobial Metals With AtomicMethyl Proply CZOI CZOI Log Reduction Log Reduction No. CMC PVADisorder^(M) Powder 0-glucan ParaBen Paraben s. aureus Ps. Aeruginosa S.Aureus Ps. Aeruginosa 5 2% 0.1% 11 13 1.4 >6 6 2% 0.5% 0.1% 0.1 0.02 1415 3.3 >6 7 2% 0.5% 0.1% 13 14 2.0 N/A 8 2% 0.5% 0.1% 0.1 14 14 2.0 N/A9 2% 0.5% 0.1% 0.20 14 14 2.0 N/A 10 2% 0.5% 0.1% 0.5 0.1 0.20 14 14 2.0>6

No. 11

A commercially available gel (glyceryl polymethacrylate) was blendedwith antimicrobial metals with atomic disorder powder to produce a gelwith a silver content of 0.1%. This gel was tested as in Nos. 5-10 andwas found to produce zones of 15 mm against both Staphylococcus aureusand Pseudomonas aeruginosa. Log reductions of 1.7 and >5 were producedagainst Staphylococcus aureus and Pseudomonas aeruginosa. This gelproduct had a greeted spectrum of activity than did mupirocin cream orointment.

No. 12

A gel coat for a urinary catheter was prepared using the formula in No.6. The coating was applied to the catheter using a dipping method. Thecoating was air dried overnight.

The dried gel coat was smooth and easy to handle. It was not tacky totouch and had excellent abrasion and adhesion properties. Uponrewetting, the surface became extremely slippery indicating excellentlubricious properties.

A zone of inhibition test was performed against Pseudomonas aeruginosausing an inoculum as prepared in No. 1. The inoculum (0.1 mL) was spreadover the surface of Mueller-Hinton agar in a Petri plate. The catheterwas cut into 1″ segments which were laid on their side into the middleof the Petri plate. Petri plates were incubated for 24 h and then thezone of inhibitions was measured.

In all cases, zones of inhibition were generated that ranged from 7-10mm. This indicates that getting drying and rehydrating had no negativeeffect on the antimicrobial activity of the gel coat.

1. A method for treating tissue, comprising: forming a free-standingpowder comprising at least one antimicrobial nanocrystalline metal withatomic disorder; and injecting the free-standing powder intherapeutically effective amounts to the tissue which is to be treated.2-22. (canceled)
 23. A method for treating tissue, comprising; forming afree-standing powder comprising at least one nanocrystalline metal; andapplying the free-standing powder in therapeutically-effective amountsto the tissue which is to be treated, wherein the tissue comprises atleast on e tissue selected from the group consisting of endocardiumtissue, pericardium tissue, bone tissue, and joint tissue. 24-30.(canceled)
 31. A method for treating tissue, comprising: forming asolution at a site remote from the tissue which is to be treated bydissolving in water at least one nanocrystalline metal; and applying thesolution in therapeutically effective amounts to the tissue which is tobe treated, wherein the tissue comprises at least one tissue selectedfrom the group consisting of endocardium tissue, pericardium tissue,bone tissue, and joint tissue. 32-46. (canceled)
 47. The method of claim1, wherein the at least nanocrystalline metal comprises nanocrystallinesilver.
 48. The method of claim 23, wherein the at least nanocrystallinemetal comprises nanocrystalline silver.
 49. The method of claim 31,wherein the at least nanocrystalline metal comprises nanocrystallinesilver.
 50. The method of claim 49, wherein the nanocrystalline silveris atomically disordered.
 51. The method of claim 31, wherein the atleast one nanocrystalline metal is atomically disordered.
 52. The methodof claim 31, wherein the solution is applied by passing it through acatheter to the tissue which is to be treated.
 53. The method of claim31, wherein the solution is applied by injection into the tissue whichis to be treated.
 54. The method of claim 53, wherein injection iseffected by the use of a needle.
 55. The method of claim 53, whereininjection is needle-less injection.
 56. The method of claim 31, whereinthe solution is applied by injection into a body cavity so as to contactthe tissue which is to be treated.